Method for Mosquito Control

ABSTRACT

A formulation and method for insect control is provided in the form of insecticide carrying insects which can be introduced in a population to thereby control the insect population. The formulation may include artificially generated adult insect carriers of a larvicide in which the larvicide has minimal impact on the adult insect and which larvicide affects juvenile survival or interferes with metamorphosis of juvenile insects to adulthood. The insects may be either male or female and may include mosquitoes.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.61/477,781, filed Apr. 21, 2011, herein incorporated by reference.

FIELD OF THE INVENTION

The present invention relates to a method and a formulation for mosquitocontrol and, in particular, a method and a formulation for mosquitocontrol which includes, but is not limited to, using mosquitoes or otherinsects for delivering agents, e.g., insecticides such as a larvicide,to an insect population to thereby control the insect population.

BACKGROUND OF THE INVENTION

Malaria, dengue and dengue haemorrhagic fever, West Nile Virus (WNV) andother encephalites, human African trypanosomiasis (HAT), humanfilariasis, dog heartworm and other pathogens important to animals areon the increase. These diseases are transmitted via insects and, inparticular, mosquitoes. Methods for controlling mosquito populationsinclude the use of pesticides and vector control methods.

Existing insecticidal control methods rely upon field technicians, whofail to find and treat many breeding sites, which can be numerous,cryptic and inaccessible. Additional methods consist of area-widetreatment via airplane or wind-assisted dispersal from truck-mountedfoggers. Unfortunately, the latter fail to treat many breeding sites andare complicated by variable environmental conditions. Barrera et al.,“Population Dynamics of Aedes aegypti and Dengue as Influenced byWeather and Human Behavior in San Juan, Puerto Rico,” PLoS NeglectedTropical Diseases, 5:e1378, 2011, describing the effects of variousbreeding sites on disease.

Surveys of natural and artificial water containers demonstratemosquitoes and other arthropods to be highly efficient in finding,inhabiting and laying eggs in variously sized, cryptic water pools,including tree holes and gutters high above ground level.

One prior formulation or method for treating mosquito populationsincludes the use of dissemination stations which are deployed in atarget environment. The dissemination stations may be laced with apesticide, including, but not limited to, a juvenile hormone analog. Thedissemination station may include a box or other structure whichattracts female mosquitoes. The mosquitoes enter the disseminationstation, become exposed to the pesticide or hormone, and carry thathormone back to affect other mosquitoes by mating. An example of thismosquito control is described in the article by Devine et al., entitled“Using adult mosquitoes to transfer insecticides to Aedes aegypti larvalhabitats,” PNAS, vol. 106, no. 28, Jul. 14, 2009.

In tests of another dissemination station, researchers showed that malesin the wild that acquire the pesticide from a station can transfer thepesticide to females during copulation. The females receiving pesticideparticles via venereal transfer were then shown to cause a significantinhibition of emergence in larval bioassays. This was reported in thearticle by Gaugler et al, entitled “An autodissemination station for thetransfer of an insect growth regulator to mosquito oviposition sites,”Med. Vet. Entomol. 2011.

In view of continuing mosquito problems, as noted, additional tools arerequired to control mosquitoes that are important as nuisance pests anddisease vectors.

SUMMARY OF THE INVENTION

The present invention is directed to a novel, self-delivering,insecticidal formulations and delivery techniques. The formulations, inone form, are larvicide treated insects, such as male mosquitoes. Theinsecticidal formulations can control medically important mosquitoes.These medically important mosquitoes include mosquitoes having aneconomic or medical importance to animal or human health. Medicallyimportant mosquitoes include those listed in the Appendix to thisdisclosure.

One aspect of the present formulations and delivery techniques relatesto a new larvicide treatment for males, as a formulation which can beused to control a mosquito population. The formulation can be generatedby exposing adult insects, such as mosquitoes and, in particular, malemosquitoes, to a pesticide, such as a juvenile hormone which affectsjuvenile survival or interferes with metamorphosis of juvenilemosquitoes and has relatively little impact on adult mosquitoes.Advantageously, the adult insects are exposed to the pesticide in acontrolled, factory environment. The factory-reared or captured from thewild adult insects which have been exposed to the pesticide are referredto as direct treated individuals (DTI). The DTI are then released intoan environment in which one wishes to control the mosquito population.The DTI control a mosquito population by interacting with untreatedindividuals (e.g., mating), such that the pesticide, e.g., a larvicide,is communicated to other individuals (known as Indirectly TreatedIndividuals; (ITI)).

In specific further embodiments, the control method uses compounds thataffect immature/juvenile stages (eggs, larvae, pupae) more than adults.A list of larvicidal compounds is maintained at the IR-4 Public HealthPesticides Database. Examples of compounds include (1) insect growthregulators such as juvenile hormone mimics or analogs, includingmethoprene, pyriproxyfen (PPF), and (2) Microbial larvicides, such asBacillus thuringiensis and Bacillus sphaericus, herein incorporated byreference. Exemplary compounds are provided in Tables 1-3, below, in theDetailed Description section.

The present invention, in one form thereof, relates to a method forinsect control. The method includes introducing insects which carry oneor more insecticides comprising at least one larvicide, to an insectpopulation, to thereby control the insect population. In one specificembodiment, the insects are adult males and the method further includesexposing the adult male insects to a pesticide which affects juvenilesurvival or interferes with metamorphosis of juvenile insects toadulthood, and which pesticide has little impact on adult insects.

In one further, specific embodiment, the insect population is a mosquitopopulation. Further, the juvenile active insecticide (i.e. larvicide)may be within a chemical class (Table 1) or biological class (Table 2).Examples within the chemical class include insect growth regulators,such as juvenile hormone analogs or compounds which mimic juvenilehormones. For example, the larvicide may be pyriproxyfen or methoprene.Examples within the biological class include viruses, bacteria,protozoa, fungi and crustacean organisms or toxic compounds that theyproduce.

The present invention, in another form thereof, relates to a formulationfor insect control which comprises an artificially generated adultinsect carrier of a larvicide. The larvicide has minimal impact on theadult insect and the larvicide interferes with metamorphosis of juvenileinsects to adulthood. In one specific formulation, the adult insect is amale mosquito and in an alternative form, the larvicide is pyriproxyfen,methoprene and microbial larvicides, including, but not limited to,Bacillus thuringiensis and Bacillus sphaericus.

BRIEF DESCRIPTION OF THE DRAWING

The sole FIGURE is a graph showing survival of treated (black) anduntreated (white) adults, with bars showing standard deviation, inaccordance with the present invention.

DETAILED DESCRIPTION

The present invention is directed to a method and a formulation formosquito control. The formulation, in one advantageous form, islarvicide treated males. The treated males are generated from medicallyimportant adult male mosquitoes obtained via factory-rearing or capturedfrom the wild. As a demonstration of the chemical class of juvenileactive insecticides (Table 1), the adult male mosquitoes are exposed toa larvicide, such as pyriproxyfen (PPF), advantageously in a controlledlaboratory or factory environment. PPF is a juvenile hormone mimic whichinterferes with metamorphosis of juvenile mosquitoes and has relativelylittle impact on adult mosquitoes. Thus, PPF is commonly used as amosquito larvicide, but is not used as an adulticide.

The treated males are subsequently referred to as the Direct TreatedIndividuals (DTI), and this is the insecticidal formulation. The DTI arereleased into areas with indigenous conspecifics. Male mosquitoes do notblood feed or transmit disease. Accordingly, male mosquitoes provideunique advantages in the present control method as couriers of thelarvicide. The DTI interact with untreated individuals (e.g., mating),such that PPF is communicated to the other individuals to produceIndirectly Treated Individuals (ITI). The PPF is delivered by both theDTI and ITI in the wild/in the environment, into the breeding areas,where the PPF accumulates to lethal doses and acts as a larvicide. It isnoted that the PPF would impact additional mosquito species that sharethe same breeding site, providing control of additional mosquitospecies.

In an alternative control method, female mosquitoes can be used as theDTI. However, female mosquitoes blood feed and can vector disease. Theuse of female mosquitoes are applicable when the females areincapacitated prior to deployment in the environment and the femaleshave limited procreation ability, bite and vector diseases.

In a further alternative method, other larvicidal active ingredients canbe used which include, but are not limited to, compounds that affectjuvenile survival or affect immature/juvenile stages of development(eggs, larvae, pupae) more than adults. A list of larvicidal compoundsis maintained at the IR-4 Public Health Pesticides Database. Examples ofcompounds include (1) insect growth regulators such as juvenile hormonemimics or analogs, including methoprene, pyriproxyfen (PPF), and (2)Microbial larvicides, such as Bacillus thuringiensis and Bacillussphaericus. Tables 1-3 provide abridged, exemplary lists of suitablecompounds.

TABLE 1 Juvenile Active Insecticide - Chemical* AzadirachtinDiflubenzuron Methoprene Neem Oil (Azadirachta indica) NovaluronPyriproxyfen S-Methoprene S-Hydropene Temephos *A list of Public HealthPesticides is maintained at the IR-4 Public Health Pesticides Database

TABLE 2 Juvenile Active Insecticide - Biological* Ascogregarine spp.Bacillus sphaericus Bacillus thuringiensis israelensis BaculovirusesCopepoda spp. Densovirinae spp. Lagenidium giganteum Microsporida spp.Spinosad Spinosyn *A list of Public Health Pesticides is maintained atthe IR-4 Public Health Pesticides Database

TABLE 3 Public Health Pesticides from the IR-4 Database*(−)-cis-Permethrin Cyfluthrin Oil of Basil, African Blue (Ocimumkilimandscharicum × basilicum) (−)-trans-Permethrin Cyhalothrin Oil ofBasil, Dwarf Bush (Ocimum basilicum var. minimum) (+)-cis-PermethrinCyhalothrin, epimer R157836 Oil of Basil, Greek Bush (Ocimum minimum)Cyhalothrin, Total Oil of Basil, Greek Column (±)-cis,trans-Deltamethrin(Cyhalothrin-L + R157836 (Ocimum × citriodorum epimer) ‘Lesbos’)(1R)-Alpha-Pinene Cypermethrin Oil of Basil, Lemon (Ocimum americanum)(1R)-Permethrin Cyphenothrin Oil of Basil, Sweet (Ocimum basilicum)(1R)-Resmethrin DDD, o,p Oil of Basil, Thai Lemon (Ocimum × citriodorum)(1R,cis) Phenothrin DDD, other related Oil of Bay Laurel (Laurusnobilis) (1R,trans) Phenothrin DDD, p,p′ Oil of Cajeput (Melaleucaleucadendra) (1S)-Alpha-Pinene DDE Oil of Cassumunar Ginger (Zingibermontanum) (1S)-Permethrin DDE, o,p Oil of Fishpoison (Tephrosiapurpurea) (E)-Beta-Caryophyllene DDT Oil of Ginger (Zingiber officinale)1,1-dichloro-2,2-bis-(4-ethyl- DDT, o,p′ Oil of Gurjun Balsam phenyl)ethane (Dipterocarpus turbinatus balsam) 1,8-Cineole DDT, p,p′ Oil ofLemon Eucalyptus (Corymbia citriodora) 1H-Pyrazole-3-carboxamide, DDVPOil of Lemon Mint (Monarda 5-amino-1-[2,6-dichloro-4- citriodora)(trifluoromethyl)phenyl]-4- [(trifluoromethyl)sulfinyl] 1-Naphthol DDVP,other related Oil of Melaleuca (Melaleuca spp.) 1-Octen-3-ol DEET Oil ofMyrcia (Myrcia spp.) 2-(2-(p-(diisobutyl) phenoxy) Deltamethrin Oil ofNutmeg (Myristica ethoxy) ethyl dimethyl fragrans) ammonium chloride2-butyl-2-ethyl-1,3- Deltamethrin (includes Oil of Palmarosa propanediolparent Tralomethrin) (Cymbopogon martinii) Deltamethrin (isomer2-Hydroxyethyl Octyl Sulfide unspecified) Orange Oil (Citrus sinensis)2-Isopropyl-4-methyl-6- Deltamethrin, other related Oregano Oil(Origanum hydroxypyrimidine vulgare) 2-Pyrroline-3-carbonitrile, 2-Desmethyl Malathion Ortho-Phenylphenol (p-chlorophenyl)-5-hydroxy-4-oxo-5- 3,7-dimethyl-6-octen-1-ol Desulfinyl FipronilOrtho-Phenylphenol, Sodium acetate Salt 3,7-dimethyl-6-octen-1-olDesulfinylfipronil Amide Oviposition Attractant A acetate3-Phenoxybenzoic Acid Diatomaceous Earth Oviposition Attractant B4-Fluoro-3-phenoxybenzoic Diatomaceous Earth, other OvipositionAttractant C acid related Absinth Wormwood Diazinon OvipositionAttractant D (Artemisia absinthium) Absinthin Diazoxon OxymatrineAcepromazine Dibutyl Phthalate Paracress Oil (Spilanthes acmella)Acetaminophen Didecyl Dimethyl Ammonium P-Cymene Chloride AcetamipridDieldrin Penfluron Acetic Acid Diethyl Phosphate Pennyroyal Oil(American False Pennyroyal, Hedeoma pulegioides) AI3-35765 DiethylthioPhosphate Peppermint (Mentha × piperita) AI3-37220 DiflubenzuronPeppermint Oil (Mentha × piperita) Alkyl Dimethyl Benzyl Dihydro AbietylAlcohol Permethrin Ammonium Chloride (60% C14, 25% C12, 15% C16) AlkylDimethyl Benzyl Dihydro-5-heptyl-2(3H)- Permethrin, other relatedAmmonium Chloride furanone (60% C14, 30% C16, 5% C12, 5% C18) AlkylDimethylethyl Benzyl Dihydro-5-pentyl-2(3H)- Phenothrin AmmoniumChloride furanone (50% C12, 30% C14, 17% C16, 3% C18) AlkylDimethylethyl Benzyl Dimethyl Phosphate Phenothrin, other relatedAmmonium Chloride (68% C12, 32% C14) Allethrin Dimethyldithio PhosphatePicaridin Allethrin II Dimethylthio Phosphate Pine Oil (Pinus pinea =Stone Pine) Allethrins Dinotefuran Pine Oil (Pinus spp.) AllicinDipropyl Isocinchomeronate Pine Oil (Pinus sylvestris = (2, 5 isomer)Scots Pine) Dipropyl Isocinchomeronate Allyl Caproate (3, 5 isomer) PineTar Oil (Pinus spp.) Allyl Isothiocyanate Dipropylene Glycol PineneAlpha-Cypermethrin d-Limonene Piperine Alpha-lonone d-PhenothrinPiperonyl Butoxide Alpha-Pinene Dried Blood Piperonyl Butoxide,technical, other related Alpha-Terpinene d-trans-Beta-CypermethrinPirimiphos-Methyl Aluminum Phosphide Esfenvalerate PMD(p-Menthane-3,8-diol) Amitraz Ester Gum Potassium Laurate PotassiumSalts of Fatty Ammonium Bicarbonate Estragole Acids AmmoniumFluosilicate Etofenprox Potassium Sorbate Anabasine Eucalyptus Oil(Eucalyptus Prallethrin spp.) Anabsinthine Eugenol Propoxur Andiroba Oil(Carapa Eugenyl Acetate Propoxur Phenol guianensis) Andiroba Oil (CarapaExtract of Piper spp. Propoxur, other related procera) Andiroba, African(Carapa Extracts of Common Juniper Putrescent Whole Egg Solids procera)(Juniperus communis) Andiroba, American (Carapa Fenchyl AcetatePyrethrin I guianensis) Anethole Fenitrothion Pyrethrin II Anise(Pimpinella anisum) Fennel (Foeniculum vulgaris) Pyrethrins Aniseed Oil(Pimpinella Fennel Oil (Foeniculum Pyrethrins and Pyrethroids, anisum)vulgaris) manufg. Residues Atrazine Fenoxycarb Pyrethrins, other relatedAvermectin Fenthion Pyrethrum Azadirachtin Fenthion Oxon Pyrethrum MarcPyrethrum Powder other Azadirachtin A Fenthion Sulfone than PyrethrinsBacillus sphaericus Fenthion Sulfoxide Pyriproxyfen Pyrrole-2-carboxylicacid, 3- Bacillus sphaericus, serotype Ferula hermonisbromo-5-(p-chlorophenyl)-4- H-5A5B, strain 2362 cyano-Pyrrole-2-carboxylic acid, 5- Bacillus thuringiensis Ferula hermonis Oil(p-chlorophenyl)-4-cyano- israelensis (metabolite of AC 303268) Bacillusthuringiensis Finger Root Oil israelensis, serotype H-14 (Boesenbergiapandurata) Quassia Bacillus thuringiensis Fipronil Quassin israelensis,strain AM 65-52 Bacillus thuringiensis Fipronil SulfoneR-(−)-1-Octen-3-ol israelensis, strain BK, solids, spores, andinsecticidal toxins, ATCC number 35646 Bacillus thuringiensis FipronilSulfoxide Red Cedar Chips (Juniperus israelensis, strain BMP 144virginiana) Bacillus thuringiensis Fragrance Orange 418228 Resmethrinisraelensis, strain EG2215 Bacillus thuringiensis Gamma-CyhalothrinResmethrin, other related israelensis, strain IPS-78 Bacillusthuringiensis Garlic (Allium sativum) Rhodojaponin-III israelensis,strain SA3A Balsam Fir Oil (Abies Garlic Chives Oil (Allium Rose Oil(Rosa spp.) balsamea) tuberosum) Basil, Holy (Ocimum Garlic Oil (Alliumsativum) Rosemary (Rosmarinus tenuiflorum) officinalis) BendiocarbGeraniol Rosemary Oil (Rosmarinus officinalis) Benzyl Benzoate GeraniumOil (Pelargonium Rosmanol graveolens) Bergamot Oil (Citrus Glyphosate,Isopropylamine Rosmaridiphenol aurantium bergamia) Salt Beta-AlanineHexaflumuron Rosmarinic Acid Beta-Caryophyllene Hydroprene RotenoneBeta-Cyfluthrin Hydroxyethyl Octyl Sulfide, R-Pyriproxyfen other relatedBeta-Cypermethrin Imidacloprid R-Tetramethrin Beta-Cypermethrin ([(1R)-Imidacloprid Guanidine Rue Oil (Ruta chalepensis) 1a(S*),3a] isomer)Beta-Cypermethrin ([(1R)- 1a(S*),3b] isomer) Imidacloprid Olefin RyaniaBeta-Cypermethrin ([(1S)- Imidacloprid Olefinic- Ryanodine 1a((R*),3a]isomer) Guanidine Beta-Cypermethrin ([(1S)- Imidacloprid UreaS-(+)-1-Octen-3-ol 1a(R*),3b] isomer) Beta-Myrcene Imiprothrin SabineneBeta-Pinene Indian Privet Tree Oil (Vitex Sabinene negundo) BetulinicAcid Ionone Sage Oil (Salvia officinalis) IR3535 (Ethyl Sassafras Oil(Sassafras Bifenthrin Butylacetylaminopropionate) albidum) Billy-GoatWeed Oil Isomalathion Schoenocaubn officinale (Ageratum conyzoides)Bioallethrin = d-trans- Allethrin Isopropyl Alcohol S-CitronellolBiopermethrin Japanese Mint Oil (Mentha Sesame (Sesamum indicum)arvensis) Bioresmethrin Jasmolin I Sesame Oil (Sesamum indicum) BitterOrange Oil (Citrus Jasmolin II Sesamin aurantium) Blend of Oils: ofLemongrass, Kerosene Sesamolin of Citronella, of Orange, of Bergamot;Geraniol, lonone Alpha, Methyl Salicylate and Allylisothioc Boric AcidL-(+)-Lactic acid S-Hydroprene Borneol Lactic Acid Silica Gel BornylAcetate Lagenidium giganteum Silver Sagebrush (Artemisia cana) BromineLagenidium giganteum Silver Sagebrush Oil (California strain) (Artemisiacana) Butane Lambda-Cyhalothrin S-Methoprene Butoxy Poly PropyleneGlycol Lambda-Cyhalothrin R ester Sodium Chloride Caffeic AcidLambda-Cyhalothrin S ester Sodium Lauryl Sulfate Solvent NaphthaCamphene Lambda-Cyhalothrin total (Petroleum), Light Aromatic CamphorLauryl Sulfate Soybean Oil (Glycine max) Camphor Octanane Lavender Oil(Lavendula Spinosad angustifolia) Canada Balsam Leaves of EucalyptusSpinosyn A (Eucalyptus spp.) Carbaryl Leech Lime Oil (Citrus Spinosyn Dhystrix) Carbon Dioxide Lemon Oil (Citrus limon) Spinosyn Factor AMetabolite Carnosic Acid Licareol Spinosyn Factor D Metabolite CarvacrolLimonene S-Pyriproxyfen Caryophyllene Linalool Succinic Acid CassumunarGinger Oil Linalyl Acetate Sulfoxide (Zingiber montanum) Castor Oil(Ricinus Linseed Oil (Linum Sulfoxide, other related communis)usitatissimum) Catnip Oil (Nepeta cataria) Lonchocarpus utilis (Cubé)Sulfur Catnip Oil, Refined (Nepeta Lupinine Sulfuryl Fluoride cataria)Cedarwood Oil (Callitropsis Magnesium Phosphide Sweet Gale Oil (Myricagale) nootkatensis = Nootka Cypress, Alaska Yellow Cedarwood) CedarwoodOil (Cedrus Malabar (Cinnamomum Tangerine Oil (Citrus deodara = DeodarCedar) tamala) reticulata) Cedarwood Oil (Cedrus spp. = Malabar Oil(Cinnamomum Tansy Oil (Tanacetum True Cedars) tamala) vulgare) CedarwoodOil (Cupressus Malaoxon Tar Oils, from Distillation of funebris =Chinese Weeping Wood Tar Cypress) Cedarwood Oil (Cupressus MalathionTarragon Oil (Artemisia spp. = Cypress) dracunculus) Cedarwood Oil(Juniper and Malathion Dicarboxylic Acid Tarwood Oil (Laxostylis alata)Cypress) Cedarwood Oil (Juniperus Malic Acid tau-Fluvalinate ashei =Ashe's Juniper, Texan Cedarwood) Cedarwood Oil (Juniperus Marigold Oil(Tagetes Teflubenzuron macropoda = Pencil Cedar) minuta) Cedarwood Oil(Juniperus spp.) Matrine Temephos Cedarwood Oil (Juniperus MenthoneTemephos Sulfoxide virginiana = Eastern Redcedar, Southern Redcedar)Cedarwood Oil (Oil of Metaflumizone Terpinene Juniper Tar = Juniperusspp.) Cedarwood Oil (Thuja Metarhizium anisopliae Terpineol occidentalis= Eastern Strain F52 Spores Arborvitae) Cedarwood Oil (Thuja spp. =Methoprene Tetrachlorvinphos, Z-isomer Arborvitae) Cedarwood Oil(unspecified) Methoprene Acid Tetramethrin Cedrene Methyl AnabasineTetramethrin, other related Cedrol Methyl Bromide Theta-CypermethrinChevron 100 Neutral Oil Methyl Cinnamate Thiamethoxam Chlordane Methylcis-3-(2 2- Thujone dichlorovinyl)-2 2- dimethylcyclopropane-1-carboxylate Chlorfenapyr Methyl Eugenol Thyme (Thymus vulgaris)Chloropicrin Methyl Nonyl Ketone Thyme Oil (Thymus vulgaris)Chlorpyrifos Methyl Salicylate Thymol Cinerin I Methyl trans-3-(2 2-Timur Oil (Zanthoxylum dichlorovinyl)-2 2- alatum)dimethylcyclopropane-1- carboxylate Cinerin II Metofluthrin TralomethrinCinerins MGK 264 (N-octyl trans-3-(2,2-Dichlorovinyl)- Bicycloheptene2,2-dimethylcyclopropane Dicarboximide) carboxylic acid Cinnamon(Cinnamomum zeylanicum) Mineral Oil Trans-Alpha-lonone Cinnamon Oil(Cinnamomum zeylanicum) Mineral Oil, Petroleum TransfluthrinDistillates, Solvent Refined Light cis-3-(2,2-Dichlorovinyl)-2,2-Mixture of Citronella Oil, trans-Ocimene dimethylcyclopropane CitrusOil, Eucalyptus Oil, carboxylic acid Pine Oil cis-Deltamethrin MMF (Poly(oxy-1,2- Transpermethrin ethanediyl), alpha- isooctadecyl-omega-hydroxy) Cismethrin Mosquito Egg Pheromone trans-Resmethrincis-Permethrin Mugwort (Artemisia vulgaris) Trichlorfon Citral MugwortOil (Artemisia Triethylene Glycol vulgaris) Citric Acid Mustard Oil(Brassica spp.) Triflumuron Citronella (Cymbopogon winterianus) MyrceneTrifluralin Citronella Oil (Cymbopogon winterianus) Naled Turmeric Oil(Curcuma aromatica) Citronellal Neem Oil (Azadirachta Uniconizole-Pindica) Citronellol Nepeta cataria (Catnip) Ursolic Acid Citrus Oil(Citrus spp.) Nepetalactone Veratridine Clove (Syzygium aromaticum)Nicotine Verbena Oil (Verbena spp.) Clove Oil (Syzygium aromaticum)Nonanoic Acid Verbenone CME 13406 Nornicotine Violet Oil (Viola odorata)Coriander Oil (Coriandrum sativum) Novaluron White Pepper (Piper nigrum)Coriandrol Ocimene Wintergreen Oil (Gaultheria spp.) Coriandrum sativumOcimum × citriodorum (Thai Wood Creosote (Coriander) Lemon Basil) CornGluten Meal Ocimum × citriodorum Wood Tar ‘Lesbos’ (Greek Column Basil)Corn Oil (Zea mays ssp. Ocimum americanum Wormwood Oil (Artemisia Mays)(Lemon Basil) absinthium) Corymbia citriodora (Lemon Ocimum basilicum(Sweet Ylang-ylang Oil (Canagium Eucalyptus) Basil) odoratum) CottonseedOil (Gossypium spp.) Ocimum basilicum var. minimum (Dwarf Bush Basil)Zeta-Cypermethrin Coumaphos Ocimum kilimandscharicum × Zinc Metal Stripsbasilicum (African Blue Basil) Cryolite Ocimum minimum (Greek BushBasil) Cube Extracts (Lonchocarpus utilis) Oil of Balsam Peru (Myroxylonpereirae) *A list of Public Health Pesticides is maintained at the IR-4Public Health Pesticides Database; Version from March 2012

In yet another alternative method, the aforementioned methods can beapplied to additional susceptible arthropods, including economically andmedically important pests (including animal and human health), where onelife stage and/or sex does not cause direct damage.

In other alternative delivery techniques, the present method can beapplied using non-targeted, beneficial or non-pest arthropods thatutilize the same breeding site as the targeted arthropod. For example,the DTI could be PPF-treated arthropods that come in contact with thetargeted insect's breeding sites. As an example, Oytiscidae adults(Predaceous Diving Beetles) could be reared or field collected andtreated with PPF to become the DTI. Additional candidate insects thatcould serve as the DTI include, but are not limited to: Diptera (e.g.,Tipulidae, Chironomidae, Psychodidae, Ceratapogonidae, Cecidomyiidae,Syrphidae, Sciaridae, Stratiomyiidae, Phoridae), Coleoptera (e.g.,Staphylinidae, Scirtidae, Nitidulidae, Oytiscidae, Noteridae) andHemiptera (e.g., Pleidae, Belostomatidae, Corixidae, Notonectidae,Nepidae).

An additional benefit of the latter strategy (i.e. non-Culicid DTI) isthat the DTI may be easier to rear, larger size (allowing increasedlevels of PPF), be less affected by the PPF, or have an increasedprobability of direct contact with the breeding site of the targetedarthropod (i.e. not necessarily rely on transfer of the PPF via mating,improved location of breeding sites).

It is noted that the species of DTI would vary based upon the specificapplication, habitat and location. For example, the regulatory issuesmay be simplified if the species used for DTI were indigenous. However,it is noted that there are numerous examples of exotic arthropods beingimported for biological control. Furthermore, different DTI species maybe more/less appropriate for urban, suburban and rural environments.

Referring to the following examples for exemplary purposes only, but notto limit the scope of the invention in any way, Aedes albopictus wereused in experiments from a colony established in 2008 from Lexington,Ky. Callosobrochus maculatus were purchased from Carolina BiologicalSupply Company (Burlington, N.C.) and maintained on mung beans (Vignaradiata). Rearing and experiments were performed in ambient conditions(˜25° C.; 80% humidity). Larvae were reared in pans with ˜500 ml waterand crushed cat food (Science Diet; Hill's Pet Nutrition, Inc.). Adultswere provided with raisins as a sugar source. For line maintenance,females were blood fed by the author.

Sumilarv 0.5 G was generously provided by Sumitomo Chemical (London,UK). Liquid PPF was purchased from Pest Control Outlet (New Port Richey,Fla.). Bacillus thuringiensis subspecies israelensis technical powderwas purchased from HydroToYou (Bell, Calif.). For application, Sumilarvgranules were crushed into a fine powder and applied using abellows-type dusting apparatus (J.T. Eaton Insecticidal Duster #530;Do-ityourself Pest Control, Suwanee, Ga.). Liquid PPF was applied usinga standard squirt bottle (WalMart, Lexington, Ky.). Treated adults wereheld in individualized bags with a raisin as a sucrose source until usedin larval bioassays. Larval bioassays were performed in 3 oz. Dixie Cups(Georgia-Pacific, Atlanta, Ga.) containing ten L3 larvae, 20 ml waterand crushed cat food.

Adult treatment does not affect survival. Male and female Ae. albopictustreated with pulverized Sumilarv showed good survival in laboratoryassays, which is indistinguishable from that of untreated controlindividuals. In an initial assay, 100% survival was observed for adultsin both the Sumilarv treated (n=8 replications) and untreated controlgroups (n=2 replications) during a two-day observation period. In asecond comparison, adults were monitored for eight days. Similar to theinitial experiment, no difference was observed between the treated andcontrol groups. Specifically, a similar average longevity was observedcomparing the Sumilarv treated (6.3±2.0 days; n=4) and undusted control(7.3 days; n=1) groups. In a third experiment, treated and untreatedadults were separated by sex and monitored for eight days. Similar toprior experiments, survival was not observed to differ between thetreated and untreated groups (FIGURE).

In a separate experiment, the survival of beetles (Callosobrochusmaculatus) dusted with Sumilarv were compared to an undusted controlgroup. In both the treatment and control groups, 100% survival wasobserved during the four day experiment.

To assess the larvicidal properties of treated adults, Sumilarv dustedadults and undusted control adults were placed individually intobioassay cups with larvae. No adults eclosed from the five assay cupsreceiving a treated adult; in contrast, high levels of adult eclosionwas observed from all four control assay cups that received an untreatedadult. Chi square analysis shows the adult eclosion resulting in assaysreceiving a treated adult to be significantly reduced compared to thatin the control group (X2 (1, N=9)=12.37, p<0.0004). The bioassayexperiment was repeated in a subsequent, larger experiment, yieldingsimilar results; adult eclosion in the treated group was significantlyreduced compared to the control group (X2 (1, N=24)=13.67, p<0.0002).

A similar bioassay was used to assess the larvicidal properties oftreated beetles. Similar to the prior results, adult eclosion fromassays in the treated beetle group was significantly reduced compared tothe control group (X2 (1, N=14)=13.38, p<0.0003).

To examine an additional formulation of PPF, an identical bioassay wasperformed, but a liquid PPF solution was applied to mosquito adults,instead of Sumilarv dust. Similar to the prior results, adult eclosionin the treated group was significantly reduced compared to the controlgroup (X2 (1, N=14)=16.75, p<0.0001).

To examine an example of the biological class of juvenile activeinsecticides (Table 2) and different active ingredients, an identicalbioassay was performed, but a powder formulation of Bacillusthuringiensis subspecies israelensis technical powder was applied tomosquito adults, using the same method as the Sumilarv dust. Similar tothe prior results, no difference was observed between the longevity oftreated versus untreated adults (X2 (1, N=15)=3.2308, p>0.09). Uponexposing larvae to treat adults, eclosion was significantly reducedcompared to the control group (X2 (1, N=28)=15.328, p<0.0001).

The results demonstrate that C. maculatus and A. albopictus adults donot experience reduced survival resulting from direct treatment with theinsecticides. Specifically, the survival of treated mosquitoes andbeetles did not differ significantly from that of the untreatedconspecifics. The results are consistent with the traits required forthe proposed application of treated adults as a self-deliveringlarvicide. Treated adults must survive, disperse and find breeding sitesunder field conditions. The results of the feasibility assays reportedhere provide evidence of an advantageous method of mosquito or otherarthropod control.

Bioassays characterizing the larvicidal properties of treated adultsshow significant lethality resulting from the presence of treatedmosquitoes and beetles. Similar results were observed for multipleformulations (i.e. dust and liquid) and multiple active ingredients.Furthermore, representative examples from each of the chemical andbiological classes (Tables 1 and 2) of juvenile active insecticides havebeen demonstrated. This is also consistent with those traits requiredfor the proposed application of treated arthropods as a self-deliveringinsecticide. Specifically, treated arthropods that reach mosquitobreeding sites can be expected to impact immature mosquitoes that arepresent at the site.

It will now be clear that the present invention is directed to a novelformulation and method for treating insect populations, including, butnot limited to, mosquito populations. Unlike prior control methods thatdisseminate a pesticide using dissemination stations, followed by aninsect in the wild entering the dissemination station to become treatedwith the pesticide, the present formulation and method starts withgenerating insect carriers in an artificial controlled environment orsetting. The insect carriers can either be factory-reared or adultscaptured from the wild. Subsequently, the carriers are released into anenvironment as the control agent or formulation. Thus, the carriers,i.e. the insects with the pesticide, are the formulation for insectcontrol, whereas, in prior methods and formulations, the formulation isa treated dissemination station, not a treated insect.

One of ordinary skill in the art will recognize that the presenttreatment, which targets insect larvae, offers advantages over prior arttechniques of insect control which target adult insects. The presentmethod is a trans-generation insect control technique which targets thenext generation of insects, whereas prior techniques target the presentgeneration, i.e. adult insects. For example, García-Munguía et al.,“Transmission of Beauveria bassiana from male to female Aedes aegyptimosquitoes,” Parasites & Vectors, 4:24, 2011 is a paper published onFeb. 26, 2011 describing mosquito control of adults and, thus, the paperdescribes the killing of the present generation of insects. TheGarcía-Munguía paper describes using fungus-treated males to deliverinsecticidal fungus to adult females. The fungus shortens the femalelifespan and reduces fecundity of adult females. This type of approach(using insects to deliver an adulticide) is not particularly novel, andhas been used in several important insect species, including examplesdescribed in Baverstock et al., “Entomopathogenic fungi and insectbehaviour: from unsuspecting hosts to targeted vectors,” Biocontrol,55:89-102, 2009.

As described above, the present technique uniquely usesfactory-treatment of adult insects with a larvicide in which thelarvicide is chemical or biological in nature. No prior techniqueincludes the manufacturing of larvicidal-treated insects fortrans-generational delivery. Further, unlike prior techniques that treatadults with fungi that kills adults, the present technique merely treatsadult males with larvicidal compounds which do not kill the adult males;rather, the treatment delivers the larvicidal compounds in atrans-generational delivery to kill the next generation, i.e. larvae.

Advantages which follow from the present technique include using theadults treated with the larvicide to communicate the larvicide to otheradults through the lifespan of the initially treated adult insect. As aresult, there is an exponential effect of the present technique whichdelivers a larvicide using treated adults to transfer the treatment toother adults, rather than prior techniques which kill the adult insect.

Further, the present technique delivers the larvicide by the treatedadults into breeding sites where the larvicide can affect and killthousands of developing, immature mosquitoes. This technique is unlikeprior techniques which merely target the adults and, thus, only kill thedirectly affected adults and not thousands of developing, immaturemosquitoes, i.e. a next generation of insects.

In addition, the present technique allows for the treatment of insectbreeding sites, including cryptic, i.e. previously unknown, breedingsites which prior insect control techniques do not treat.

Further, the present technique allows one to affect insect populationsof the species of a treated insect, as well as other species which sharea common breeding site. Since the present technique uses adult insectsto deliver a larvicide to a breeding site, the present technique allowsfor the transmission of a larvicide to breeding sites which may becommon among more than one insect species. As a result, the presenttechnique can target the species of the treated insect, as well asinsects which share a common breeding site.

In addition, in contrast to adulticide methods, the present larvicidetechnique allows a pesticide to persist in a breeding site after thetreated insect has departed or died.

One additional advantage of the present method is that the agents beingdisseminated are the insects themselves, as carriers of the insecticidewhich will directly affect an insect population. Prior formulation andmethods require indirect dissemination, in which insects of a populationin the wild must first find a dissemination station, acquire anappropriate dose of the insecticide, and then return to the populationwith the larvicide of a dissemination station in order to have an affecton the insect population.

It will now be clear to one of ordinary skill in the art that thepresent formulation of pesticide carrier insects and the present methodfor controlling insect populations based on the present experiments. Forexample, if the insects have a larval stage, adult insects can be usedas carriers of larvicides which have minimal affect on the adult insect,but are lethal to the larvae, thereby controlling the insect population.

While the invention has been described in connection with numerousembodiments, it is to be understood that the specific mechanisms andtechniques which have been described are merely illustrative of theprinciples of the invention, numerous modifications may be made to themethods and apparatus described without departing from the spirit andscope of the invention.

What is claimed is:
 1. A method for insect control, comprising:introducing insects which carry one or more insecticides comprising atleast a larvicide, to an insect population, to thereby control theinsect population.
 2. The method of claim 1, wherein the insects areadult insects, and the method further comprises exposing the adultinsects to a pesticide which affects juvenile survival or interfereswith metamorphosis of juvenile insects to adulthood, and which pesticidehas little impact on adult insects.
 3. The method of claim 2, whereinthe adult insects are male insects.
 4. The method of claim 1, whereinthe insect population is a mosquito population.
 5. The method of claim2, wherein the insect population is a mosquito population.
 6. The methodof claim 3, wherein the insect population is a mosquito population. 7.The method of claim 6, wherein the adult male insects are mosquitoes. 8.The method of claim 2, wherein the pesticide is a chemical agent.
 9. Themethod of claim 8, wherein the chemical agent is selected from the groupconsisting of Azadirachtin, Diflubenzuron, Methoprene, Neem Oil(Azadirachta indica), Novaluron, Pyriproxyfen, S-Methoprene, S-Hydropeneand Temephos.
 10. The method of claim 2, wherein the pesticide is abiological agent.
 11. The method of claim 10, wherein the biologicalagent is selected from the group consisting of Ascogregarine spp.,Bacillus sphaericus, Bacillus thuringiensis israelensis, Baculoviruses,Copepoda spp., Densovirinae spp., Lagenidium giganteum, Microsporidaspp., Spinosad and Spinosyn.
 12. The method of claim 1, wherein thelarvicide is a juvenile hormone analog or a compound which mimics ajuvenile hormone.
 13. The method of claim 12, wherein the larvicide ispyriproxyfen.
 14. The method of claim 2, wherein exposing the adultinsects to a pesticide comprises exposing the adult males to thepesticide in a controlled environment to produce directly treatedindividuals and the introducing insects comprises introducing thedirectly treated individuals into the insect population.
 15. The methodof claim 14, further comprising allowing the direct treated individualsto interact with untreated individuals of the insect population, towhich the directly treated individual insects have been introduced, tothereby produce indirectly treated individuals.
 16. The method of claim15, wherein both the directly treated individuals and the indirectlytreated individuals control the insect population by delivering thepesticide to the insect population.
 17. The method of claim 1, whereinthe insects carrying the pesticide are female.
 18. A formulation forinsect control, said formulation comprising: an artificially generatedadult insect carrier of a larvicide, wherein said larvicide has minimalimpact on the adult insect and which larvicide affects juvenile survivalor interferes with metamorphosis of juvenile insects to adulthood. 19.The formulation of claim 18, wherein the adult insect is a malemosquito.
 20. The formulation of claim 18, wherein the adult insect is afemale mosquito.
 21. The formulation of claim 18, wherein the larvicideis a chemical agent.
 22. The formulation of claim 21, wherein thechemical agent is selected from the group consisting of Azadirachtin,Diflubenzuron, Methoprene, Neem Oil (Azadirachta indica), Novaluron,Pyriproxyfen, S-Methoprene, S-Hydropene and Temephos.
 23. Theformulation of claim 18, wherein the larvicide is a biological agent.24. The formulation of claim 23, wherein the biological agent isselected from the group consisting of Ascogregarine spp., Bacillussphaericus, Bacillus thuringiensis israelensis, Baculoviruses, Copepodaspp., Densovirinae spp., Lagenidium giganteum, Microsporida spp.,Spinosad and Spinosyn.